Nanopore-Based Biomimetic ssDNA Reporters for Multiplexed Detection of Meat Adulteration

Meat adulteration detection is crucial for ensuring food safety, protecting consumer rights, and maintaining market integrity. However, the current methods face challenges in achieving multiplexed detection through efficient signal conversion and output. This study introduces a nanopore-based approach for the simultaneous detection of multiple meat adulteration. Leveraging the interaction between DADA dipeptide and vancomycin, we designed a series of biomimetic ssDNA reporters with DADA-Van tags for multiplexed signal output. These ssDNA reporters can generate highly distinctive current blockage signals, which can be discriminated simultaneously by the current waveform. Combined with PCR-based strand displacement, ssDNA reporters can be released from magnetic beads in the presence of the target gene and are analyzed using alpha-hemolysin (alpha-HL) nanopores for multiplexed signal output. The signal frequency for each target gene has a linear relationship with the specific concentration range (for duck: 1 x 10-3 similar to 10 nmol/L, for pork: 1 x 10-2 similar to 10 nmol/L, and for chicken: 1 x 10-3 similar to 10 nmol/L). The limits of detection are as follows: 0.418 pmol/L for duck, 4.473 pmol/L for pork, and 0.531 pmol/L for chicken. This method effectively enables the simultaneous detection of adulterated chicken, pork, and duck meat in lamb samples and holds potential for broad applications in ensuring the authenticity and safety of meat products.